WNK1 mediates M-CSF-induced macropinocytosis to enforce macrophage lineage fidelity

Alissa J. Trzeciak; Zong-Lin Liu; Mohamed Gatie; Adam S. Krebs; Waleska Saitz Rojas; Anya J. O'Neal; Ann Baako; Zhaoquan Wang; Justin Nelson; Isabella C. Miranda; Jazib Uddin; Allie Lipshutz; Jian Xie; Chou-Long Huang; Pedro H.V. Saavedra; Anna-Katerina Hadjantonakis; Michael Overholtzer; Michael S. Glickman; Arohan R. Subramanya; Thomas Vierbuchen; Jon Iker Etchegaray; Christopher D Lucas; Christopher N. Parkhurst; Justin S.A. Perry

doi:10.1038/s41467-025-59901-0

WNK1 mediates M-CSF-induced macropinocytosis to enforce macrophage lineage fidelity

Alissa J. Trzeciak^*, Zong-Lin Liu, Mohamed Gatie, Adam S. Krebs, Waleska Saitz Rojas, Anya J. O'Neal, Ann Baako, Zhaoquan Wang, Justin Nelson, Isabella C. Miranda, Jazib Uddin, Allie Lipshutz, Jian Xie, Chou-Long Huang, Pedro H.V. Saavedra, Anna-Katerina Hadjantonakis, Michael Overholtzer, Michael S. Glickman, Arohan R. Subramanya, Thomas VierbuchenJon Iker Etchegaray, Christopher D Lucas, Christopher N. Parkhurst, Justin S.A. Perry^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

Tissue-resident macrophages (TRMs) are critical for mammalian organismal development and homeostasis. Here, we report that with-no-lysine 1 (WNK1) controls myeloid progenitor fate, with Csf1riCre-mediated Wnk1 deletion in mice (WNK1-deficient mice) resulting in loss of TRMs and perinatal mortality. Mechanistically, macrophage-colony stimulating factor (M-CSF) induced continuous macropinocytosis in mouse and human myeloid progenitors and monocytes, which was lost with WNK1-deficiency or WNK kinase activity inhibition and boosted with WNK1 overexpression. Absence of WNK1, inhibition of WNK kinase activity, or disruption of M-CSF stimulated macropinocytosis blocked mouse and human progenitor and monocyte differentiation into macrophages, instead skewing progenitor differentiation into neutrophils. Treatment with phorbol-12-myristate-13-acetate (PMA) rescued macropinocytosis but not macrophage differentiation of WNK-inhibited progenitors, suggesting that M-CSF-stimulated macropinocytosis-induced activation of WNK1 is required for macrophage differentiation. Finally, M-CSF-stimulated macropinocytosis triggered WNK1 nuclear translocation and concomitant increased protein expression of interferon regulatory factor (IRF)8, whereas inhibition of macropinocytosis or WNK kinase activity suppressed IRF8 expression. Thus, we discovered a role of macropinocytosis, via WNK1 activity, for myeloid cell lineage commitment during development and homeostasis.

Original language	English
Article number	4945
Number of pages	19
Journal	Nature Communications
Volume	16
DOIs	https://doi.org/10.1038/s41467-025-59901-0
Publication status	Published - 28 May 2025

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WNK1 mediates M-CSF-induced macropinocytosis to enforce macrophage lineage fidelityFinal published version, 5.09 MBLicence: Creative Commons: Attribution (CC-BY)

Dissecting macrophage regulation of lung epithelial regeneration
Lucas, C. (Principal Investigator)
Medical Research Council
1/08/23 → 31/07/28
Project: Research
Resolving innate inflammatory responses to tissue injury and apoptotic cell clearance to develop novel therapeutic strategies for pulmonary diseases
Wood, W. (Principal Investigator), Bain, C. (Co-investigator), Jenkins, S. (Co-investigator), Lucas, C. (Co-investigator) & Rossi, A. (Co-investigator)
Medical Research Council
1/06/22 → 31/05/27
Project: Research

Cite this

Trzeciak, A. J., Liu, Z.-L., Gatie, M., Krebs, A. S., Saitz Rojas, W., O'Neal, A. J., Baako, A., Wang, Z., Nelson, J., Miranda, I. C., Uddin, J., Lipshutz, A., Xie, J., Huang, C.-L., Saavedra, P. H. V., Hadjantonakis, A.-K., Overholtzer, M., Glickman, M. S., Subramanya, A. R., ... Perry, J. S. A. (2025). WNK1 mediates M-CSF-induced macropinocytosis to enforce macrophage lineage fidelity. Nature Communications, 16, Article 4945 . https://doi.org/10.1038/s41467-025-59901-0

@article{671c2cde9d0940828ab9923a7f2f67dc,

title = "WNK1 mediates M-CSF-induced macropinocytosis to enforce macrophage lineage fidelity",

abstract = "Tissue-resident macrophages (TRMs) are critical for mammalian organismal development and homeostasis. Here, we report that with-no-lysine 1 (WNK1) controls myeloid progenitor fate, with Csf1riCre-mediated Wnk1 deletion in mice (WNK1-deficient mice) resulting in loss of TRMs and perinatal mortality. Mechanistically, macrophage-colony stimulating factor (M-CSF) induced continuous macropinocytosis in mouse and human myeloid progenitors and monocytes, which was lost with WNK1-deficiency or WNK kinase activity inhibition and boosted with WNK1 overexpression. Absence of WNK1, inhibition of WNK kinase activity, or disruption of M-CSF stimulated macropinocytosis blocked mouse and human progenitor and monocyte differentiation into macrophages, instead skewing progenitor differentiation into neutrophils. Treatment with phorbol-12-myristate-13-acetate (PMA) rescued macropinocytosis but not macrophage differentiation of WNK-inhibited progenitors, suggesting that M-CSF-stimulated macropinocytosis-induced activation of WNK1 is required for macrophage differentiation. Finally, M-CSF-stimulated macropinocytosis triggered WNK1 nuclear translocation and concomitant increased protein expression of interferon regulatory factor (IRF)8, whereas inhibition of macropinocytosis or WNK kinase activity suppressed IRF8 expression. Thus, we discovered a role of macropinocytosis, via WNK1 activity, for myeloid cell lineage commitment during development and homeostasis.",

author = "Trzeciak, {Alissa J.} and Zong-Lin Liu and Mohamed Gatie and Krebs, {Adam S.} and {Saitz Rojas}, Waleska and O'Neal, {Anya J.} and Ann Baako and Zhaoquan Wang and Justin Nelson and Miranda, {Isabella C.} and Jazib Uddin and Allie Lipshutz and Jian Xie and Chou-Long Huang and Saavedra, {Pedro H.V.} and Anna-Katerina Hadjantonakis and Michael Overholtzer and Glickman, {Michael S.} and Subramanya, {Arohan R.} and Thomas Vierbuchen and Etchegaray, {Jon Iker} and Lucas, {Christopher D} and Parkhurst, {Christopher N.} and Perry, {Justin S.A.}",

year = "2025",

month = may,

day = "28",

doi = "10.1038/s41467-025-59901-0",

language = "English",

volume = "16",

journal = "Nature Communications",

issn = "2041-1723",

publisher = "Nature Publishing Group",

}

Trzeciak, AJ, Liu, Z-L, Gatie, M, Krebs, AS, Saitz Rojas, W, O'Neal, AJ, Baako, A, Wang, Z, Nelson, J, Miranda, IC, Uddin, J, Lipshutz, A, Xie, J, Huang, C-L, Saavedra, PHV, Hadjantonakis, A-K, Overholtzer, M, Glickman, MS, Subramanya, AR, Vierbuchen, T, Etchegaray, JI, Lucas, CD, Parkhurst, CN & Perry, JSA 2025, 'WNK1 mediates M-CSF-induced macropinocytosis to enforce macrophage lineage fidelity', Nature Communications, vol. 16, 4945 . https://doi.org/10.1038/s41467-025-59901-0

TY - JOUR

T1 - WNK1 mediates M-CSF-induced macropinocytosis to enforce macrophage lineage fidelity

AU - Trzeciak, Alissa J.

AU - Liu, Zong-Lin

AU - Gatie, Mohamed

AU - Krebs, Adam S.

AU - Saitz Rojas, Waleska

AU - O'Neal, Anya J.

AU - Baako, Ann

AU - Wang, Zhaoquan

AU - Nelson, Justin

AU - Miranda, Isabella C.

AU - Uddin, Jazib

AU - Lipshutz, Allie

AU - Xie, Jian

AU - Huang, Chou-Long

AU - Saavedra, Pedro H.V.

AU - Hadjantonakis, Anna-Katerina

AU - Overholtzer, Michael

AU - Glickman, Michael S.

AU - Subramanya, Arohan R.

AU - Vierbuchen, Thomas

AU - Etchegaray, Jon Iker

AU - Lucas, Christopher D

AU - Parkhurst, Christopher N.

AU - Perry, Justin S.A.

PY - 2025/5/28

Y1 - 2025/5/28

N2 - Tissue-resident macrophages (TRMs) are critical for mammalian organismal development and homeostasis. Here, we report that with-no-lysine 1 (WNK1) controls myeloid progenitor fate, with Csf1riCre-mediated Wnk1 deletion in mice (WNK1-deficient mice) resulting in loss of TRMs and perinatal mortality. Mechanistically, macrophage-colony stimulating factor (M-CSF) induced continuous macropinocytosis in mouse and human myeloid progenitors and monocytes, which was lost with WNK1-deficiency or WNK kinase activity inhibition and boosted with WNK1 overexpression. Absence of WNK1, inhibition of WNK kinase activity, or disruption of M-CSF stimulated macropinocytosis blocked mouse and human progenitor and monocyte differentiation into macrophages, instead skewing progenitor differentiation into neutrophils. Treatment with phorbol-12-myristate-13-acetate (PMA) rescued macropinocytosis but not macrophage differentiation of WNK-inhibited progenitors, suggesting that M-CSF-stimulated macropinocytosis-induced activation of WNK1 is required for macrophage differentiation. Finally, M-CSF-stimulated macropinocytosis triggered WNK1 nuclear translocation and concomitant increased protein expression of interferon regulatory factor (IRF)8, whereas inhibition of macropinocytosis or WNK kinase activity suppressed IRF8 expression. Thus, we discovered a role of macropinocytosis, via WNK1 activity, for myeloid cell lineage commitment during development and homeostasis.

AB - Tissue-resident macrophages (TRMs) are critical for mammalian organismal development and homeostasis. Here, we report that with-no-lysine 1 (WNK1) controls myeloid progenitor fate, with Csf1riCre-mediated Wnk1 deletion in mice (WNK1-deficient mice) resulting in loss of TRMs and perinatal mortality. Mechanistically, macrophage-colony stimulating factor (M-CSF) induced continuous macropinocytosis in mouse and human myeloid progenitors and monocytes, which was lost with WNK1-deficiency or WNK kinase activity inhibition and boosted with WNK1 overexpression. Absence of WNK1, inhibition of WNK kinase activity, or disruption of M-CSF stimulated macropinocytosis blocked mouse and human progenitor and monocyte differentiation into macrophages, instead skewing progenitor differentiation into neutrophils. Treatment with phorbol-12-myristate-13-acetate (PMA) rescued macropinocytosis but not macrophage differentiation of WNK-inhibited progenitors, suggesting that M-CSF-stimulated macropinocytosis-induced activation of WNK1 is required for macrophage differentiation. Finally, M-CSF-stimulated macropinocytosis triggered WNK1 nuclear translocation and concomitant increased protein expression of interferon regulatory factor (IRF)8, whereas inhibition of macropinocytosis or WNK kinase activity suppressed IRF8 expression. Thus, we discovered a role of macropinocytosis, via WNK1 activity, for myeloid cell lineage commitment during development and homeostasis.

U2 - 10.1038/s41467-025-59901-0

DO - 10.1038/s41467-025-59901-0

M3 - Article

SN - 2041-1723

VL - 16

JO - Nature Communications

JF - Nature Communications

M1 - 4945

ER -

WNK1 mediates M-CSF-induced macropinocytosis to enforce macrophage lineage fidelity

Abstract

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Dissecting macrophage regulation of lung epithelial regeneration

Resolving innate inflammatory responses to tissue injury and apoptotic cell clearance to develop novel therapeutic strategies for pulmonary diseases

Cite this